用于细胞外囊泡检测的功能分子集成纳米界面

IF 3.5
Hirobumi Sunayama, Yuya Matsui, Toshifumi Takeuchi
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引用次数: 0

摘要

通过模板聚合和多步后修饰制备了用于细胞外囊泡检测的功能分子集成聚合物纳米空腔。制备了一种具有叔胺基的多功能聚合物,用于与模板二氧化硅纳米颗粒(NPs)和可聚合的甲基丙烯酸基相互作用,并与500 nm的二氧化硅纳米颗粒络合。该配合物单分散固定在羧基和引入到镀金玻璃基板上的Br基上。此外,表面引发的2-甲基丙烯酰氧乙基磷酸胆碱原子转移自由基聚合形成生物相容性聚合物层。去除二氧化硅NPs后,使用三(2-羧基乙基)膦裂解二硫键以去除多功能聚合物组分,产生巯基暴露的纳米空腔。通过改变荧光染料和非荧光染料的混合比例来控制功能化程度。通过引入荧光染料和硝基三乙酸片段,然后引入his标记的蛋白G和CD9蛋白的抗体,制备了多功能纳米腔,产生ev传感纳米界面。该荧光信号接口以浓度依赖的方式响应ev,估计检测限为0.1 fM。当参比抗体被引入时,这些反应没有被观察到,证实了空腔选择性功能化。该方法可用于生命科学领域中多功能纳米界面的制备。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Functional Molecule-Integrated Nano-Interfaces for Detection of Extracellular Vesicles

Functional Molecule-Integrated Nano-Interfaces for Detection of Extracellular Vesicles

Functional molecule-integrated polymer nanocavities for the detection of extracellular vesicles (EVs) are prepared via template polymerization and multistep post-modification. A multifunctional polymer bearing a tertiary amino group for interaction with template silica nanoparticles (NPs) and a polymerizable methacryl group is prepared and complexed with 500-nm silica NPs. The complex is monodispersedly immobilized on the carboxyl group and Br group introduced onto the Au-coated glass substrate. Additionally, surface-initiated atom transfer radical polymerization of 2-methacryloyloxyethyl phosphorylcholine is conducted to form a biocompatible polymer layer. After the silica NPs are removed, the disulfide bond is cleaved using tris(2-carboxyethyl)phosphine to remove the multifunctional polymer components, yielding a thiol group-exposed nanocavity. The degree of functionalization is controlled by changing the mixing ratio of the fluorescent and non-fluorescent dyes. A multifunctionalized nanocavity is prepared by introducing a fluorescent dye and nitrilotriacetic acid moiety, followed by His-tagged protein G and an antibody for the CD9 protein, yielding EV-sensing nano-interfaces. This fluorescent signaling interface responds to EVs in a concentration-dependent manner, with an estimated limit of detection of 0.1 fM. These responses are not observed when the reference antibody is introduced, confirming cavity-selective functionalization. The proposed method is useful for fabricating multifunctional nano-interfaces in the field of life sciences.

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